摘要
目的构建一种以猴痘病毒(monkeypox virus,MPXV)优势保护性抗原为基础的多表位疫苗。方法从NCBI网站获取9种MPXV蛋白的GenBank登录ID及其氨基酸序列;通过ABCpred、NetCTL 1.2和IEDB的MHC-I Immunogenicity模块分别预测9种MPXV蛋白的B、CTL及Th细胞(HTL)表位;采用IEDB的MHC I Binding模块预测各CTL表位与特定MHC-I类分子的亲和力;使用VaxiJen v2.0、ToxinPred、AllerTOP v.2.0分别预测所得表位的抗原性、毒性及致敏性,从中筛选出具有最高抗原性但无细胞毒性和致敏性的表位,其中,CTL表位选择IC_(50)值<30 nmol/L者,HTL表位选择IFN-γ预测值最好者;将上述各表位与RS09、PADRE、Pam2Cys三种佐剂随机连接并添加6×H(组蛋白)尾巴以构建多表位疫苗,从各连接组合中筛选出具有最高抗原性但无细胞毒性和致敏性的组合;通过ExPASy的Protparam与ProtScale模块分析其理化性质;通过PSIPRED预测其二级结构;使用I-TASSER进行多表位疫苗的三级结构模拟,然后使用ModeRefiner和GalaxyRefiner将模拟出的三级结构进行优化;使用ZDOCK serves将优化后的多表位疫苗与TLR2、TLR4受体进行分子对接处理,以观察其相互作用情况与对接位点;采用C-ImmSim行免疫模拟对疫苗的免疫效果进行评估。结果该MPXV疫苗包括9个B细胞表位、23个CTL细胞表位和9个HTL表位,组合方式为RS09-PADRE-B-Pam2Cys-HTL-CTL-6×H,其抗原性得分为0.6828,共包含675个氨基酸残基,相对分子质量(M_r)为74431.30,理论pI值9.36,分子式为C_(3473)H_(5267)N_(867)O_(920)S_(17),热稳定性好,亲水性强;二级结构中α螺旋占46.82%,β折叠占18.37%,无规则卷曲占34.81%;可与TLR2、TLR4分子稳定结合,还可诱导较强的体液与细胞免疫应答。结论运用免疫信息学方法成功构建一种含多个优势抗原表位的MPXV疫苗,可能对猴痘病毒感染有预防作用。
Objective To develop a multi-epitope vaccine based on the dominant protective antigen of monkeypox virus(MPXV).Methods GenBank login ID and amino acid sequences of 9 MPXV proteins were obtained from NCBI website.The B,CTL and Th cell(HTL)epitopes of 9 MPXV proteins were predicted by the MHC-I Immunogenicity modules of ABCpred,NetCTL 1.2 and IEDB,respectively;The MHC I Binding module of IEDB was used to predict the affinity between CTL epitopes and specific MHC-I molecules.VaxiJen v2.0,ToxinPred and AllerTOP v.2.0 were used to predict the antigenicity,toxicity and allergenicity of these epitopes.Epitopes with the highest antigenicity but no cytotoxicity and allergenicity were selected.Among them,the CTL epitopes with IC_(50)<30 nmol/L and HTL epitopes with the highest IFN-γpredictive value were selected.Allthe epitopes were randomly connected with RS09,PADRE and Pam2Cys adjuvants and a 6×H(histone)tail was added to construct the multi-epitope vaccine.The combination with the highest antigenicity but no cytotoxicity and allergenicity was selected;physical and chemical properties were analyzed using Protparam and ProtScale modules of ExPASy;its secondary structure was predicted by PSIPRED;its tertiary structure was simulated using I-TASSER and then the simulated tertiary structure was optimized using ModeRefiner and GalaxyRefiner;the molecular docking of optimized multi-epitope vaccines with TLR2 and TLR4 receptors was performed by ZDOCK serves,and their interactions and docking sites were analyzed.The immunological effect of the vaccine was evaluated by C-ImmSim immunization simulation.Results The combination mode of the proposed MPXV vaccine,consisting of 9 B-cell epitopes,23 CTL epitopes and 9 HTL epitopes,was RS09-PADRE-B-Pam2Cys-HTL-CTL-6 H,and its antigenality score was 0.6828,containing 675 amino acid residues,with a relative molecular mass(Mr)of 74431.30 and a theoretical pI value of 9.36.Its molecular formula was C_(3473)H_(5267)N_(867)O_(920)S_(17),withing good thermal stability and strong hydrophilicity
作者
宋浩然
辛继莹
王亚
孙艳丽
SONG Haoran;XIN Jiying;WANG Ya;SUN Yanli(School of Medical Laboratory,Weifang Medical University,Weifang 261053,Shandong,China)
出处
《中国病原生物学杂志》
CSCD
北大核心
2024年第8期869-874,879,共7页
Journal of Pathogen Biology
基金
山东省自然科学基金面上项目(No.ZR2020MH379)
2023年潍坊医学院大学生创新创业训练计划项目(No.X2023623)
2023年潍坊医学院大学生创新创业训练计划项目(No.X2023630)。
